In Vivo Cross-Linking MS Reveals Conservation in OmpA Linkage to Different Classes of β-Lactamase Enzymes

J Am Soc Mass Spectrom. 2020 Feb 5;31(2):190-195. doi: 10.1021/jasms.9b00021. Epub 2019 Nov 21.

Abstract

Molecular interactions between two different classes of β-lactamase enzymes and outer membrane protein A (OmpA) were studied by in vivo chemical cross-linking of a multi-drug-resistant strain of Acinetobacter baumannii AB5075. Class A β-lactamase blaGES-11 and Class D β-lactamase Oxa23, responsible for hydrolysis of different types of β-lactam antibiotics, were found to be cross-linked to similar lysine sites of the periplasmic domain of outer membrane protein OmpA, despite low sequence homology between the two enzymes. The findings from in vivo XL-MS suggest that the interacting surfaces between both β-lactamase enzymes and OmpA are conserved during molecular evolution, and the OmpA C-terminus domain serves an important function of anchoring different types of β-lactamase enzymes in the periplasmic space.

Keywords: AB5075; OmpA; Oxa23; blaGES-11; in vivo cross-linking.

MeSH terms

  • Acinetobacter baumannii / chemistry
  • Acinetobacter baumannii / metabolism*
  • Amino Acid Sequence
  • Bacterial Outer Membrane Proteins / chemistry
  • Bacterial Outer Membrane Proteins / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Conserved Sequence
  • Cross-Linking Reagents / chemistry
  • Mass Spectrometry
  • Models, Molecular
  • Protein Conformation
  • Protein Interaction Domains and Motifs
  • Protein Interaction Maps
  • beta-Lactamases / chemistry
  • beta-Lactamases / metabolism*

Substances

  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Cross-Linking Reagents
  • OMPA outer membrane proteins
  • beta-Lactamases